title: Quality control of astrocyte-directed Cre transgenic mice: the benefits of a direct link between loss of gene expression and reporter activation.
abstract: Cre recombinase activity for cell-type restricted deletion of floxed target genes (i.e., flanked by Cre recognition loxP-sites) is often measured by separate matings with recombination-activated reporter gene mice. Using a floxed Gja1 (Cx43) allele, we demonstrate the benefits of a direct link between reporter gene expression and target gene deletion to overcome critical limitations of the Cre/loxP system. The widely used human glial fibrillary acidic protein (hGFAP)-Cre transgene exhibits variable recombination activity and requires postexperimental validation. Such quality control is essential to correlate the extent of Cre-mediated Gja1 ablation with phenotypical alterations and to maintain the activity status of hGFAP-Cre in transgenic mouse colonies. We present several strategies to control for the fidelity of hGFAP-Cre mediated recombination. (c) 2008 Wiley-Liss, Inc.
authors.affiliation: Institute of Genetics, Division of Molecular Genetics, University of Bonn, D-53117 Bonn, Germany.
authors.forename: Robert Pascal
authors.initials: RP
authors.lastname: Requardt
authors.forename: Lech
authors.initials: L
authors.lastname: Kaczmarczyk
authors.forename: Pavel
authors.initials: P
authors.lastname: Dublin
authors.forename: Anke
authors.initials: A
authors.lastname: Wallraff-Beck
authors.forename: Thomas
authors.initials: T
authors.lastname: Mikeska
authors.forename: Joachim
authors.initials: J
authors.lastname: Degen
authors.forename: Andreas
authors.initials: A
authors.lastname: Waha
authors.forename: Christian
authors.initials: C
authors.lastname: Steinhäuser
authors.forename: Klaus
authors.initials: K
authors.lastname: Willecke
authors.forename: Martin
authors.initials: M
authors.lastname: Theis
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000502:physiology; D017630:Connexins / Q000235:genetics; D005260:Female; D017353:Gene Deletion; D015870:Gene Expression; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005904:Glial Fibrillary Acidic Protein / Q000235:genetics; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D008297:Male; D008745:Methylation; D051379:Mice; D008822:Mice, Transgenic; D011401:Promoter Regions, Genetic / Q000502:physiology; D011786:Quality Control
chemical_list: D017630:Connexins; D005904:Glial Fibrillary Acidic Protein